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GNPS - Malpighiaceae plants (137 species)

ABSTRACT: Analyses of 136 Malpighiaceae species (most of them from Brazil, most of them leaves). Dried plant material were extracted, separately, with EtOH80% and EtOAc. Positive and Negative ionization mode analises. LC-MS/MS performed in an UltiMate 3000 UPLC system (Thermo Scientific) using a Kinetex 1.7 mm C18 reversed phase UHPLC column (50 X 2.1 mm) and Maxis Impact Q-TOF mass spectrometer (Bruker Daltonics) equipped with ESI source.

INSTRUMENT(S): impact HD|MS:1002667

ORGANISM(S): Dicella Macroptera (ncbitaxon:1027124) Niedenzuella Poeppigiana Dicella Bracteosa (ncbitaxon:1027122) Stigmaphyllon Ciliatum (ncbitaxon:1027211) Byrsonima Spicata (ncbitaxon:883765) Peixotoa Hispidula Stigmaphyllon Alternifolium Bunchosia Montana Byrsonima Sericea Bunchosia Pallescens Stigmaphyllon Urenifolium Bronwenia Megaptera Niedenzuella Lucida Stigmaphyllon Convolvulifolium Tetrapterys Cardiophylla Peixotoa Tomentosa Hiraea Cuiabensis Stigmaphyllon Bonariense Banisteriopsis Vernoniifolia Banisteriopsis Adenopoda (ncbitaxon:1027063) Barnebya Harleyi Diplopterys Pubipetala (ncbitaxon:1027129) Byrsonima Ligustrifolia Heteropterys Leschenaultiana (ncbitaxon:1027148) Stigmaphyllon Caatingicola Stigmaphyllon Puberulum Heteropterys Pauciflora (ncbitaxon:259665) Callaeum Psilophyllum (ncbitaxon:1027114) Banisteriopsis Parvglandula Byrsonima Intermedia Amorimia Concinna Stigmaphyllon Sinuatum Camarea Affinis Thryallis Latifolia (ncbitaxon:151883) Acridocarpus Smeathmannii (ncbitaxon:217129) Heteropterys Oberdanii Ptilochaeta Densiflora Heteropterys Aenea Banisteriopsis Anisandra Byrsonima Incarnata Sphedamnocarpus Angolensis (ncbitaxon:1027204) Peixotoa Glabra (ncbitaxon:151863) Mascagnia Conformis Niedenzuella Multiglandulosa Stigmaphyllon Cavernulosum Tetrapterys Microphylla (ncbitaxon:151881) Stigmaphyllon Palmatum Aenigmatanthera Lasiandra (ncbitaxon:212253) Peixotoa Spinensis Stigmaphyllon Macropodum Callaeum Antifebrile (ncbitaxon:1027110) Peixotoa Reticulata (ncbitaxon:1027199) Stigmaphyllon Acuminatum Banisteriopsis Malifolia Tetrapterys Ambigua (ncbitaxon:1027214) Heteropterys Tomentosa Heteropterys Intermedia Byrsonima Crassifolia (ncbitaxon:4270) Banisteriopsis Megaphylla Hiraea Hatschbachii Stigmaphyllon Gayanum Stigmaphyllon Salzmannii Stigmaphyllon Harleyi Hiraea Reclinata (ncbitaxon:1125931) Stigmaphyllon Saxicola Verrucularina Glaucophylla (ncbitaxon:151891) Heteropterys Campestris Amorimia Septentrionalis Banisteriopsis Argyrophylla (ncbitaxon:1027065) Heteropterys Brunnea Tetrapterys Mucronata Heladena Multiflora (ncbitaxon:151830) Heteropterys Coleoptera Amorimia Pubiflora Heteropterys Hatschbachii Banisteriopsis Muricata (ncbitaxon:1027072) Heteropterys Bicolor (ncbitaxon:259657) Stigmaphyllon Angustilobum Heteropterys Brachiata (ncbitaxon:1027142) Byrsonima Coccolobifolia (ncbitaxon:1027102) Peixotoa Cordistipula (ncbitaxon:1027196) Stigmaphyllon Hispidum Niedenzuella Stannea (ncbitaxon:151855) Camarea Ericoides Amorimia Andersonii Janusia Mediterranea (ncbitaxon:151842) Mcvaughia Bahiana (ncbitaxon:151857) Blepharandra Heteropetala (ncbitaxon:151797) Hiptage Benghalensis (ncbitaxon:151834) Stigmaphyllon Auriculatum Janusia Janusioides (ncbitaxon:1027163) Stigmaphyllon Blanchetii Acmanthera Latifolia (ncbitaxon:151786) Bunchosia Glandulifera (ncbitaxon:1027090) Byrsonima Verbascifolia Banisteriopsis Quadriglandula Malpighia Mexicana (ncbitaxon:1027172) Stigmaphyllon Hatschbachii Banisteriopsis Laevifolia (ncbitaxon:1027070) Niedenzuella Sericea (ncbitaxon:1027194) Niedenzuella Acutifolia (ncbitaxon:1027193) Heteropterys Chrysophylla Heteropterys Byrsonimifolia (ncbitaxon:259658) Heteropterys Eglandulosa Aspicarpa Harleyi (ncbitaxon:1027057) Janusia Schwannioides Ptilochaeta Bahiensis (ncbitaxon:151869) Galphimia Brasiliensis (ncbitaxon:1027132) Heteropterys Leona (ncbitaxon:176641) Stigmaphyllon Paralias (ncbitaxon:151877) Alicia Anisopetala Diacidia Aracaensis Carolus Chasei Christianella Multiglandulosa (ncbitaxon:1027120) Stigmaphyllon Vitifolium Banisteriopsis Membranifolia Gaudichaudia Albida (ncbitaxon:151825) Tetrapterys Phlomoides (ncbitaxon:1027218) Burdachia Duckei Mascagnia Sepium (ncbitaxon:151854) Stigmaphyllon Lalandianum Blepharandra Hypoleuca (ncbitaxon:1027080) Banisteriopsis Stellaris Amorimia Coriacea Tetrapterys Ramiflora Hiraea Restingae Stigmaphyllon Occidentale Stigmaphyllon Tomentosum Verrucularia Piresii Ectopopterys Soejartoi (ncbitaxon:151817) Banisteriopsis Harleyi (ncbitaxon:1027069) Banisteriopsis Variabilis Heteropterys Umbellata Banisteriopsis Campestris Tetrapterys Schiedeana (ncbitaxon:1027220) Janusia Occhionii Tetrapterys Xylosteifolia

SUBMITTER: Pieter Dorrestein Vanderlan da Silva Bolzani

PROVIDER: MSV000085119 | GNPS | Wed Mar 18 19:27:00 GMT 2020

REPOSITORIES: GNPS

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Project description:Sample preparation was conducted using an adaptation of our SimPLIT workflow10. Four cell pellets corresponding to four different colorectal cancer cell lines were lysed in a buffer containing 1% sodium deoxycholate (SDC, Merck, Cat. No. 30970), 100 mM triethylammonium bicarbonate (TEAB, Merck, Cat. No. T7408), 10% isopropanol and 50 mM NaCl, freshly supplemented with 5 mM TCEP (Thermo, Bond-breaker Cat. No 77720), 10 mM iodoacetamide (IAA, Merck, Cat. No. I1149), universal nuclease 1:2000 vol/vol (Pierce, Cat. No. 88700) and Halt protease and phosphatase inhibitor cocktail (Thermo, Cat. No. 78442, 100X) with 5 min of bath sonication. Protein concentration was measured with the Quick Start Bradford protein assay (Bio-Rad, Cat. No. 5000205). Aliquots of 30 μg of total protein from each cell line were transferred in a 96-well plate according to the 58-plex design. An E. coli protein extract (Bio-Rad, Cat. No. 1632110) was added at 1 μg or 0.5 μg across the human cell lysates. The plate was dried and the samples for the arginine-specific cleavage set (TrypR) were reconstituted in 25 μL (TMT11 subset) or 12.5 μL (TMT18 subset) of 100 mM TEAB buffer followed by the addition of 10 μL TMT11 (stock 20 μg/μL, Thermo Fischer Scientific, Cat. No. A34808) or 5 μL TMTpro18 (stock 25 μg/μL, Thermo Fischer Scientific, Cat. No. A52045), and 1h reaction at room temperature for protein-level TMT labelling. The TMT labelled samples were SpeedVac dried, and all samples (labelled and unlabelled) were reconstituted in 20 μL of 100 mM TEAB buffer. Proteins were digested overnight at room temperature by the addition of 2 μL LysC (Thermo Fischer Scientific, Cat. No. 90307) or Trypsin (Thermo Fischer Scientific, Cat. No. 90059) stock solution of 500 ng/μL in 0.1% formic acid (final enzyme 50 ng/μL, 1:30) for the LysC and TrypR sets respectively. The peptides were then fully labelled by another addition of 10 μL TMT11 or 5 μL TMTpro18. The reaction was quenched after 1h with the addition of 2 μL 5% hydroxylamine solution and premix samples were prepared by pooling 0.5 μL from each sample into 100 μL of 0.1% TFA for each sub-plex separately. In each premix, SDC was precipitated with addition of formic acid at 2% and centrifugation at 20,000 × g for 5 minutes. Supernatants were transferred into clean vials for LC-MS analysis. Following evaluation of the premix runs, samples were pooled into four sub-plexes, diluted with 100 μL of HPLC water, heated at 95 °C for 5 min, cooled down and heated for another 5 minutes. Lastly, each pool was acidified with formic acid at 2%, the precipitated SDC was removed by centrifugation and supernatants were SpeedVac dried.High-pH Reversed-phase Peptide FractionationAll four sub-plexes were pooled in 100 μL of 0.1% (v/v) ammonium hydroxide and the final peptide pool was fractionated with high pH Reversed-Phase chromatography using the XBridge C18 column (2.1 × 150 mm, 3.5 μm, Waters) on an UltiMate 3000 HPLC system over a 1% gradient in 35 min. Mobile phase A was 0.1% (v/v) ammonium hydroxide and mobile phase B was 0.1% ammonium hydroxide (v/v) in acetonitrile. Fractions were collected every 30 sec and eventually pooled into 45 samples for LC-MS analysis.

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GNPS - Malpighiaceae plants (137 species)

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